Two lysosomal hydrolases from Acanthamoeba castellanii have been isolated from ameba growth medium and highly purified. N-acetyl-Beta-hexosaminidase was purified approximately 3500 fold over the growth medium. The activity applied to SDS polyacrylamide gels ran as a single band of 54,000 molecular weight. On sizing columns the activity eluted with an apparent molecular weight of 110,000, suggesting that the enzyme is a dimer. Beta-glucosidase was purified 6000 fold over the growth medium. The apparent molecular weight from both sizing columns and polyacrylamide gel electrophoresis was 84,000. On PAGE one major and several minor bands were found. Preparative gel electrophoresis was performed and the 84,000 MW band eluted for injection into rabbits. Polyclonal antibodies to both purified hydrolyses have been obtained.